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PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The involvement of LTB4 in the recruitment of neutrophils in combination with other chemoattractants to inflamed joints was also shown (65, 66).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Our data suggest that the synergistic effect of LTB4 and CXCL12 is specific for CXCR4-induced chemotaxis since LTB4 has no effect on CXCR5-mediated migration of VAL-wt.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Nevertheless, we cannot fully exclude that CXCL12 scavenging by ACKR3 creates microgradients, which somehow modify CXCR4/CXCL12 signaling on ACKR3 negative cells.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Further studies are required to describe the molecular mechanism that supports the release of LTB4 upon ACKR3/CXCR4 stimulation.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Using a diffuse xenograft model we showed that spreading of VAL cells into tissues, such as brain, bone marrow and spleen, requires ACKR3 expression (32).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Given the poor prognosis of DLBCL patients with brain infiltrations (67, 68), understanding the underlying mechanism is of great clinical relevance.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Here we report that ACKR3 is required for DLBCL to escape from local tumors to move to draining lymph nodes resembling the spread of malignant cells from lumps or extra nodal sites.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Hence, targeting ACKR3-mediated tumor dissemination is appealing.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Our data showing the involvement of the LTB4/BLT1R axis in VAL cell migration, could open a way for the development of novel therapeutic approaches in the treatment of DLBCL, potentially in combination with inhibitors of ACKR3 (69–72).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
VAL and DOHH2 cells were cultured in RPMI-1640 supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS), 1% Penicillin/Streptomycin (P/S), 1% GlutaMAX.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
1% NEAA, 1% Sodium-Pyruvate and 50 µM β-mercaptoethanol all from Thermo Fisher.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Primary murine lymphatic endothelial cells (mLEC) were isolated from lymph nodes of C57BL/6 mice and cultured in αMEM (Thermo Fisher) supplemented with 10% heat-inactivated FBS, 1% P/S. Cells were cultured in humidified air at 37° C with 5% CO2.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Freshly harvested lymph nodes were harvested from C57BL/6 mice, and incubated in digestion mixture containing RPMI-1640, 0.25 mg/mL Liberase TL (Roche, Cat: 5401020001), 200 Kunits/mL DNAse I (Sigma-Aldrich, Cat: 10104159001) at 37° C for 1h with occasional shaking.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Single cell suspensions were strained using a 70 µm cell strainer (Corning), centrifuged and resuspended in αMEM.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were plated on 6-well plates coated with 10 µg/mL PureCol (Collagen, Sigma-Aldrich) and human 10 µg/mL plasma Fibronectin (Sigma-Aldrich).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
After 3 hours non-adhering cells were removed.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
5-7 days later, when >80% confluence was achieved, cells were detached using Accutase (Biological Industries).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Under these conditions blood endothelial cells die and the remaining cells are composed of fibroblastic reticular cells (FRCs) (~40%) and LECs.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
LECs were purified by staining with APC-conjugated anti-mouse CD31 (Biolegend, clone 390) and positively selected using anti-APC microbeads according to manufacturer’s instructions (Miltenyi Biotec).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
ACKR3 and the C-terminus deficient variant ACKR3-ΔC (6) were cloned into CDH-IF1-MCS-COP-GFP vector and nucleofected (Lonza) into CFP VAL-ko (32) giving rise to VAL-wt rescue and VAL-ACKR3-ΔC.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The transfected constructs expressed GFP after a T2A cleavage sequence, which was used for clonal selection by flow cytometry and single cell sorting (FACS-ARIA).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
CRISPR/Cas-9 ribonucleoproteins (crRNPs) targeting blt1r gene were delivered to VAL-wt cells by transfection with NEON nucleofector (Invitrogen, MA, USA) at 1’600 V, 10 ms, 3 pulses, using the provided buffer R. 10 cells were used with the 10-μl tip transfection.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The crRNAs (/AltR1/rCrArU rGrArG rUrCrU rArGrA rCrCrG rCrUrC rArCrG rUrUrU rUrArG rArGrC rUrArU rGrCrU/AltR2/”), tracrRNAs (IDT, USA) and HiFi Cas9 Nuclease V3 (IDT, USA) transfection was performed as indicated in the manufacturer’s instruction (IDT, USA).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Deletion of blt1r was confirmed by genomic sequencing of the locus.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
For flow cytometry cells were suspended in MACS buffer (phosphate buffered saline (PBS), 2% FBS, 2mM EDTA) and stained for 15 minutes on ice with the indicated antibodies.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
For chemokine uptake, 50 nM of fluorescently labelled chemokine (42, 73) was added to cells for 45 min at 37°C.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Mean fluorescence intensity (MFI) of the fluorophore were determined by flow cytometry (Fortessa or FACSCanto (BD) and FlowJo software.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Lymphoma cell migration was measured in transmigration assays using 5 µm transwell inserts.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
mLECs were seeded on polycarbonate inserts coated with 10 µg/mL fibronectin and 10 µg/mL collagen.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Monolayers of endothelial cells were grown by adding medium to the upper compartment, leaving the lower compartment empty.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
For LTB4-induced migration lymphoma cells were embedded in a collagen matrix formed by 1.6 mg/mL PureCol, 0.36% PBS supplemented with 0.36% FBS, 0.036% P/S, 1.5 µg/mL recombinant human ICAM-1/CD54 Fc chimera (R&D systems) at 4° C. To induce collagen polymerization the temperature was slowly raised over 45 min to 37°C.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Then medium was added on the upper compartment of the transwell inserts for 24 h before the experiment.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
When indicated, embedded cells were pretreated with 1 µM BIIL315 (OpnMe-Boehringer Ingelheim) for one hour at 37°C.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The medium of the upper compartment was then replaced with fresh medium containing 1 µM BIIL315, which was also present in the lower compartment during the experiment.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Lymphoma cells were pretreated with 10 µM AMD3100 (Sigma-Aldrich) for one hour at 37° C and transwell migration was performed in the presence of the same concentration of the inhibitor in the upper and lower compartment.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were treated with 2 µg/mL pertussis toxin (PTX) (Sigma-Aldrich) for two hours at 37°C.,
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
washed and added to inserts coated with endothelial cells without toxin.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Migration was stimulated with chemoattractants at the indicated concentrations in the lower compartment.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were allowed to migrate to the lower chamber for 8 hours.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
After a brief centrifugation (1 min, 250 x g) of the inserts, cells were collected by centrifugation and resuspended in 100 µL of MACS buffer.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
For cell number determination cells were counted for 50 sec by flow cytometry.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were resuspended at 10 cells mL in PBS and incubated for 30 minutes on ice.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were centrifuged at 350g for 3 minutes, re-suspended at 10 cells mL in plain RPMI and incubated for 30 minutes at 37°C.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Secretion of LTB4 was stopped by centrifugation at 4° C (350g for 3 minutes) and supernatants collected.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
LTB4 ELISA was performed as indicated in the manufacturer’s instruction (Enzo Life Sciences).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Lymphoma cell migration in 3D chemotaxis were performed using µ-Slide Chemotaxis (Ibidi).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were embedded in a collagen matrix as described above.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
To optimize and standardize the architecture of the collagen fibers we slowly induced polymerization by precooling and extending the warming rate from 4°C (liquid) to 37°C (solid) of the collagen matrix (74) (75).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The IBD chambers were loaded at 4°C and slowly warmed up to room temperature (20 min) before placing into a 37°C cell incubator.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Total warming time to 37°C was about 40 min.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The condition allows the formation of thin fibers with homogenous diameter, length and density and prevents the formation of fibers in hierarchical structures (74) (75).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
After polymerization complete medium (65 µL) was added on both side-reservoirs.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells embedded in the collagen matrix in the µ-Slide chamber were incubated in a humidified environment at 37°C and 5% CO2 for one day.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
After 24 hours 15 µL of 400 nM CXCL12 were added to one of the reservoirs and migration observed for 6 hours by time-lapse video microscopy (ImageXpress Molecular Devices) at 2 minutes or 20 seconds time intervals between frames.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The localized mouse xenograft model was described previously (32).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
8–10-week-old mice were injected subcutaneously with 10 VAL or DOHH2 cells in PBS, or only PBS for control groups.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
All in vivo experiments were performed following the rules of the Swiss Federal Veterinary Office guidelines and authorized by the Animal Studies Committee of Cantonal Veterinary.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
A day before culling 50 µL of Evans Blue (2 µg/mL) were injected subcutaneously on the flank close to the xenograft do visualize lymphatic vessels.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Localized tumors and draining axillary lymph node were surgically removed and disintegrated with 150 µm Sefar Nitex filters and cells processed for flow cytometry analysis as described (76).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells of the disintegrated lymph node were resuspended in 150 µL of MACS buffer and analyzed by flow cytometry.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Tumors and lymph nodes were collected and fixed with a solution of 4% paraformaldehyde and washed with PBS, 1% FBS and 0.05% NaN3.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The organs were embedded in low melting agarose (Sigma-Aldrich) and 50-100 µm sections were prepared with a Leica VT1200S Vibratome.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Images were acquired with a confocal microscope (Leica SP5) and analyzed with Imaris software (Bitplane).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
For each condition, five biological replicates of cells were separately cultured, treated and sorted.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
DLBCL VAL cells were stained with 5 µg/mL anti-human ACKR3 (mAb 11G8).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
at 4°C for 15 min.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Cells were washed twice with MACS Buffer and sorted (5x10 cells) for ACKR3 positivity or negativity in 500 µL of RNA-later (Sigma-Aldrich).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
RNA-later lysates were centrifuged at 2500g x 5 minutes.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The Zymo column RNA Isolation kit was used for RNA extraction (Zymo).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Transcripts were processed for RNAseq with E7765L NEBNext Ultra II Directional RNA Library Prep, Sample Purification Beads and E7490L NEBNext Poly(A) mRNA Magnetic Isolation Module.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
Poly-A RNA sequencing was performed with Illumina NextSeq500.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
RNA quantification was performed using Salmon v1.4.0 with hg38 transcriptome reference (77).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The expression levels of the transcripts were then compared in the R package DESeq2 v1.28.0 (78).
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The datasets presented in this study can be found in online repositories.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The names of the repository/repositories and accession number(s) can be found here https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE169144.
PMC9878562
ACKR3 promotes CXCL12/CXCR4-mediated cell-to-cell-induced lymphoma migration through LTB4 production
The animal study was reviewed and approved by Swiss Federal Veterinary Office guidelines and authorized by the Animal Studies Committee of Cantonal Veterinary.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
The treatment of B cell malignancies has dramatically changed with the introduction of immunotherapy, especially chimeric antigen receptor T (CAR-T) cell therapy.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
However, only limited efficacy is observed in acute myeloid leukaemia (AML).
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
In the study, We detected CD123 and CLL-1 expression on leukaemia cells from Relapsed/Refractory AML (R/R AML) patients.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Then, we constructed anti-CD123 CAR and CLL-1 CAR with different co-stimulation domains (CD28 or 4-1BB) and detected their anti-AML effects.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
To increase the efficacy of CAR-T cell therapy, we tested different strategies, including application of combined checkpoint inhibitors and histone deacetylase inhibitors (HDACi) in vivo and in vitro.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
We found CD123 and CLL-1 were highly expressed on AML cells.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
The proportions of T cell subsets and NK cells involved in anti-tumour or anti-inflammation processes in AML patients significantly decreased when compared with healthy donors.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Both CD123 CAR and CLL-1 CAR displayed specific anti-AML effects in vitro.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
To improve the lysis effects of CAR-T cells, we combined CAR-T cell therapy with different agents.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
PD-1/PD-L1 antibodies only slightly improved the potency of CAR-T cell therapy (CD123 CAR-T 60.92% ± 2.9087% vs. 65.43% ± 2.1893%, 60.92% ± 2.9087% vs. 67.43% ± 3.4973%; 37.37% ± 3.908% vs. 41.89% ± 5.1568%, 37.37% ± 3.908% vs. 42.84% ± 4.2635%).
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
However, one HDACi (valproic acid [VPA]) significantly improved CAR-T cell potency against AML cells (CLL-1 CAR-T 34.97% ± 0.3051% vs. 88.167% ± 1.5327%, p < 0.0001; CD123 CAR-T 26.87% ± 2.7010% vs. 82.56% ± 3.086%, p < 0.0001 in MV411; CLL-1 CAR-T 78.77% ± 1.2061% vs. 93.743% ± 1.2333%, p < 0.0001; CD123 CAR-T 64.10% ...
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Combination therapy prolonged the overall survival of mice when compared with single CD123 CAR-T cell therapy (median survival: 180 days vs. unfollowed).
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
A possible mechanism is that activated CD8+T cells upregulate natural-killer group 2 member D (NKG2D), and VPA upregulates NKG2D ligand expression in AML cells, contributing to NKG2D-mediated cytotoxicity of CAR-T cells against tumour cells.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
In conclusion, CD123 and CLL-1 are promising targets for AML CAR-T cell therapy.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
A combination of VPA pre-treatment and CAR-T against AML exhibits synergic effects.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
In recent years, preclinical studies of chimeric antigen receptor T (CAR-T) to treat acute myeloid leukemia (AML) have yielded promising results.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Several tumor antigens targeted to AML have been explored, including CD33, CD123, C-type lectin-like molecule-1 (CLL-1), CD44v6, and CD7.1–4 Among them, CD123 and CLL-1 remain the most promising targets for AML.2 5 However, the short persistence of CAR-T cells and immune escape may result in a relapse of AML.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Natural-killer group 2 member D (NKG2D) is normally expressed in all natural killer (NK), NK T cells (NKT), CD8 T cells, and subsets of γδ T cells.6 NKG2D recognizes a family of major histocompatibility complex (MHC) I chain-related molecules A and B and a family of six UL16-binding proteins 1-6 (ULBP1–6), which are wi...
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
HDACi can influence tumor immunogenicity, the microenvironment, and the functional activity of specific immune cells.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Moreover, HDACi was reported to upregulate NKG2DL in AML.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Therefore, it is likely that a combination of CAR-T cell therapy with agents involved in T cell activation could be a novel potency to treat AML by activating T cells.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
In the present study, we constructed anti-CD123 CAR and CLL-1 CAR, which displayed specific anti-AML effects in vivo and in vitro.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
To improve the therapeutic effect and overcome immune escape, we tested the combined CAR-T cell therapy with HDACi and programmed cell death protein-1 (PD-1) or its ligand PD-L1 antibodies to treat AML.
PMC10391797
Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia
Interestingly, one HDACi, valproic acid (VPA), could significantly synergize with CAR-T cell therapy to improve antitumor activity.